Microplastics, Alien Species and Amphibian Male-specific Predation in River Otter Diet (lutra Lutra). a Study of Two Populations in the Ticino Valley (north Italy) and Sila Massif (south Italy).

Abstract Distribution and diet of Eurasian otter (Lutra lutra) was studied in two different Italian areas: in the North (Ticino River) and in the South (streams from Sila Massif). The Ticino population derives from a reintroduction project performed by the two Park Autorities in the 90?, while the Sila Massif population are probably the descendent of relictus populations. The Ticino population is characterised by few founders coming from European reproduction centers. The release, in 1997, of a pair of otters Lutra lutra, from the breeding enclosure centre of Cameri ?Bosco Vedro? (Novara) and after the escape during a flood of two adult and a cub in 1993 from the breeding centre ?La Fagiana? (Magenta), create a little vital otter nucleous in the valley. The survey performed in 2010 provideed a small range of otter activity (7 km near the breeding centre of Cameri), in 2012 an otter sighting has been recorded and in 2013 an individual has been found dead in Southern stretch of the Ticino River near Pavia, about 30 km downstream the release site. Seven years after the last monitoring, a new survey was performed along all the Ticino River. Status and distribution of the Ticino?s otters showed a surprising evolution of its distribution. 101 feces samples were recorded in 16 stations on the 32 survayed (50%). Positive stations covered near the whole watercourse from Lake Maggiore to Pavia for a length of 100 km of river. In otter diet, the allochtonous european catfish (S. glanis) has been recorded in Italy for the first time. The high incidence of this species in the otter diet reflects its demographic explosion in Ticino River and suggests river otter such potential natural predator to control the exponential diffusion of this alien fish. To quantify the presence of S. glanis in the otter diet, morphometric equations were developed basing on 28 S. glanis speciemens, coming from the same area. Regression equations relate the known fish mass to skeletrical element dimension. Two diagnostic elements were selected starting from those found in otter stools: vertebrae and a basicranial bone. A second unexpected result found in otter feces from Ticino River was that, unfortunality, for the first time, spectroscopy analysis performed on some unknown remains in otter stool showed theirs correlation with microplastics. Probably the mustelid ingested these particles not by directly ingestion but through fish preyed. Among the multiple human pressures on aquatic ecosystems, the accumulation of plastic debris is an increasing environmental problem in marine and freshwater system. The transfer of microplastic pollution across trophic levels has been proven in marine ecosystems such between mussels and crabs. The similarity between ecological niches and feeding strategies for species in marine and freshwater environments would suggest that trophic transfer should also occur in freshwater ecosystems. 5 Considering toxicological problems related to additives release and adsorbed pollutants, this result rises a new treat for otter conservation, and certainly requires further studies. South Italy represent for Italian otter population the main core. Although in 20th century the mustelid suffered an important constriction of its range also here, recent works showed an expansion in this zone as elsewhere. The south Mediterranean area is a delicate ecological environment characterized by extreme seasonal variations in water flow with dry and harsh period in summer and torrential floods usually occuring in autumn and spring. This conditions could make the trophic resources and the river accessibility for otters highly fluctuating. Strategies for facing fluctuations in habitat accessibility and food resources are essential for the lifestyle of otter which is metabolically costly and strictly linked to aquatic habitats. We analysed the southern Italian range on 8 streams flowing from the Sila massif, and we found a stable and positive trend of the otter presence. 357 otter stools were found from 2014 utill 2017 principally in Savuto, Amato, Lese and Neto rivers. Diet analysis confirmed an increase of trophic diversity from northern to southern Europe with a high proportion of amphibian consumption. Usually diet analyses focuses on fish preys, while amphibians are lees detailed with only one or few categories. Little number of studies classified amphibians to the species level. The high presence of amphibians in the present study induced us to perform an accurate species and genus analysis. Osteological identification of amphibian remains allowed to ascertain otter predation on seven out of the 11 species recorded for the study area. Particularly interesting was the discovery of newts in otters? feces for the presence of the powerful neurotoxin in skin secretion of these urodelans. Furthermore, osteological analysis provided a male-biased predation by otter on Anurans. Stool analysis provided further interesting insights on the feeding behavior of otter. By the analysis of unknown remains performed with Attenuated Total Reflectance (ATR-FT-IR) spectroscopy, the 61% of these materials showed peaks typical of protein and high correlations with amphibian eggs. According to other studies, results of this thesis showed that Italian otter populations are following a positive trend of expansion both in the North than in the South. The diet results highlighted the plastic feeding behavior of otter, through which otters feed on the most accessible prey: alien species in the North and amphibians, even newts, in the South, with a male specific predation. The presence of amphibian eggs in otter stools may derive from the ingestion of females before spawning.